LOCUS       V042621                 7609 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042621
VERSION     V042621
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             874..1590
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1642..3099
                     /label="TRIM58(NM_015431)"
                     /note="TRIM58(NM_015431)"
                     /gene="TRIM58"
     polyA_signal    3176..3400
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3446..3874
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3888..4217
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4284..5078
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5252..5373
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5422..5438)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5446..5462
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5470..5500)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5515..5536
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5824..6409)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6580..7440)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7441..7545)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catggcctgg gcgccgcccg gggagcggct gcgcgaggat
     1681 gcgcggtgcc cggtgtgcct ggatttcctg caggagccgg tcagcgtgga ctgcggccac
     1741 agcttctgcc tcaggtgcat ctccgagttc tgcgagaagt cggacggcgc gcagggcggc
     1801 gtctacgcct gtccgcagtg ccggggcccc ttccggccct cgggctttcg ccccaaccgg
     1861 cagctggcgg gcctggtgga gagcgtgcgg cggctggggt tgggcgcggg gcccggggcg
     1921 cggcgatgcg cgcggcacgg cgaggacctg agccgcttct gcgaggagga cgaggcggcg
     1981 ctgtgctggg tgtgcgacgc cggccccgag cacaggacgc accgcacggc gccgctgcag
     2041 gaggccgccg gcagctacca ggtaaagctc cagatggctc tggaacttat gaggaaagag
     2101 ttggaggacg ccttgactca ggaggccaac gtggggaaaa agactgtcat ttggaaggag
     2161 aaagtggaaa tgcagaggca gcgcttcaga ttggagtttg agaagcatcg tggctttctg
     2221 gcccaggagg agcaacggca gctgaggcgg ctggaggcgg aggagcgagc gacgctgcag
     2281 agactgcggg agagcaagag ccggctggtc cagcagagca aggccctgaa ggagctggcg
     2341 gatgagctgc aggagaggtg ccagcgcccg gccctgggtc tgctggaggg tgtgagagga
     2401 gtcctgagca gaagtaaggc tgtcacaagg ctggaagcag agaacatccc catggaactg
     2461 aagacagcat gctgcatccc tgggaggagg gagctcttaa ggaagttcca agtggatgta
     2521 aagctggatc ccgccacggc gcacccgagt ctgctcttga ccgccgacct gcgcagtgtg
     2581 caggatggag aaccatggag ggatgtcccc aacaaccctg agcgatttga cacatggccc
     2641 tgcatcctgg gtttgcagag cttctcatca gggaggcatt actgggaggt tctggtggga
     2701 gaaggagcag agtggggttt aggggtctgt caagacacac tgccaagaaa gggggaaacc
     2761 acgccatctc ctgagaatgg ggtctgggcc ctgtggctgc tgaaagggaa tgagtacatg
     2821 gtccttgcct ccccatcagt gcctcttctc caactggaaa gtcctcgctg cattgggatt
     2881 ttcttggact atgaagccgg tgaaatttca ttctacaatg tcacagatgg atcttatatc
     2941 tacacattca accaactctt ctctggtctt cttcggcctt actttttcat ctgtgatgca
     3001 actcctctta tcttgccacc cacaacaata gcagggtcag gaaattgggc atccagggat
     3061 catttagatc ctgcttctga tgtaagagat gatcatctct aaggatcctg atatccagca
     3121 cagtggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     3181 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3241 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3301 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3361 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3421 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3481 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3541 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3601 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3661 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3721 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3781 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3841 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3901 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3961 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4021 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4081 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4141 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4201 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4261 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4321 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4381 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4441 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4501 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4561 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4621 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4681 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4741 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4801 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4861 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4921 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4981 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5041 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5101 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5161 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5221 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5281 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5341 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5401 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5461 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5521 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5581 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5641 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5701 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5761 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5821 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5881 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5941 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6001 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6061 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6121 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6181 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6241 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6301 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6361 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6421 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6481 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6541 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6601 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6661 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6721 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6781 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6841 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6901 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6961 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7021 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7081 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7141 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7201 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7261 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7321 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7381 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7441 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7501 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7561 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggt
//
